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Title: Directed Network Wiring Identifies a Key Protein Interaction in Embryonic Stem Cell Differentiation

Authors:
; ; ; ; ; ; ; ; ; ; ;  [1];  [2];  [2]
  1. (Mount Sinai Hospital)
  2. (
Publication Date:
Research Org.:
Argonne National Lab. (ANL), Argonne, IL (United States). Advanced Photon Source (APS)
Sponsoring Org.:
USDOE Office of Science (SC)
OSTI Identifier:
1148600
Resource Type:
Journal Article
Resource Relation:
Journal Name: Mol. Cell; Journal Volume: 54; Journal Issue: (6) ; 06, 2014
Country of Publication:
United States
Language:
ENGLISH

Citation Formats

Yasui, Norihisa, Findlay, Greg M., Gish, Gerald D., Hsiung, Marilyn S., Huang, Jin, Tucholska, Monika, Taylor, Lorne, Smith, Louis, Boldridge, W. Clifford, Koide, Akiko, Pawson, Tony, Koide, Shohei, Toronto), and UC). Directed Network Wiring Identifies a Key Protein Interaction in Embryonic Stem Cell Differentiation. United States: N. p., 2014. Web. doi:10.1016/j.molcel.2014.05.002.
Yasui, Norihisa, Findlay, Greg M., Gish, Gerald D., Hsiung, Marilyn S., Huang, Jin, Tucholska, Monika, Taylor, Lorne, Smith, Louis, Boldridge, W. Clifford, Koide, Akiko, Pawson, Tony, Koide, Shohei, Toronto), & UC). Directed Network Wiring Identifies a Key Protein Interaction in Embryonic Stem Cell Differentiation. United States. doi:10.1016/j.molcel.2014.05.002.
Yasui, Norihisa, Findlay, Greg M., Gish, Gerald D., Hsiung, Marilyn S., Huang, Jin, Tucholska, Monika, Taylor, Lorne, Smith, Louis, Boldridge, W. Clifford, Koide, Akiko, Pawson, Tony, Koide, Shohei, Toronto), and UC). Mon . "Directed Network Wiring Identifies a Key Protein Interaction in Embryonic Stem Cell Differentiation". United States. doi:10.1016/j.molcel.2014.05.002.
@article{osti_1148600,
title = {Directed Network Wiring Identifies a Key Protein Interaction in Embryonic Stem Cell Differentiation},
author = {Yasui, Norihisa and Findlay, Greg M. and Gish, Gerald D. and Hsiung, Marilyn S. and Huang, Jin and Tucholska, Monika and Taylor, Lorne and Smith, Louis and Boldridge, W. Clifford and Koide, Akiko and Pawson, Tony and Koide, Shohei and Toronto) and UC)},
abstractNote = {},
doi = {10.1016/j.molcel.2014.05.002},
journal = {Mol. Cell},
number = (6) ; 06, 2014,
volume = 54,
place = {United States},
year = {Mon Jul 28 00:00:00 EDT 2014},
month = {Mon Jul 28 00:00:00 EDT 2014}
}
  • Embryonic stem cells (ESCs) often display high rates of apoptosis and spontaneous differentiation in routine culture, thus bring the proliferation of these cells highly inefficient. Moreover, little is known about the factors that are indispensable for sustaining self-renewal. To surmount these issues, we established transgenic mES cell lines expressing the enhanced green fluorescent protein (EGFP) under the control of the Rex1 promoter which is a key regulator of pluripotency in ES cells. In addition, we provided a simplified and improved protocol to derive transgenic mESCs from single cell. Finally, we showed that embryoid body (EB) development was faster than adherentmore » differentiation in terms of differentiation ratio by real-time tracking of the EGFP expression. Therefore, these cell lines can be tracked and selected both in vitro and in vivo and should be invaluable for studying the factors that are indispensable for maintaining pluripotency.« less
  • Research on stem-cell transplantation has indicated that the success of transplantation largely depends on synchronizing donor cells into the G0/G1 phase. In this study, we investigated the profile of embryonic stem (ES) cell synchronization and its effect on the formation of embryonic bodies (EBs) using cell culture with serum deprivation. The D3 cell line of ES cells was used, and parameters such as cell proliferation and activity, EB formation, and expression of stage-specific embryonic antigen-1 and Oct-4 were investigated. Results showed that the percentage of G0/G1 stage in serum deprivation culture is significantly higher than that in culture with serummore » supplementation. Synchronized ES cells can reenter the normal cell cycle successfully after serum supply. EBs formed from synchronized ES cells have higher totipotency capability to differentiate into functional neuronal cells than EBs formed from unsynchronized ES cells. Our study provides a method for ES treatment before cell transplantation that possibly helps to decrease the rate of cell death after transplantation.« less
  • Highlights: •Down-regulating FUT9 and ST3Gal4 expression blocks L1-induced neuronal differentiation of ESCs. •Up-regulating FUT9 and ST3Gal4 expression in L1-ESCs depends on the activation of PLCγ. •L1 promotes ESCs to differentiate into neuron through regulating cell surface glycosylation. -- Abstract: Cell recognition molecule L1 (CD171) plays an important role in neuronal survival, migration, differentiation, neurite outgrowth, myelination, synaptic plasticity and regeneration after injury. Our previous study has demonstrated that overexpressing L1 enhances cell survival and proliferation of mouse embryonic stem cells (ESCs) through promoting the expression of FUT9 and ST3Gal4, which upregulates cell surface sialylation and fucosylation. In the present study,more » we examined whether sialylation and fucosylation are involved in ESC differentiation through L1 signaling. RNA interference analysis showed that L1 enhanced differentiation of ESCs into neurons through the upregulation of FUT9 and ST3Gal4. Furthermore, blocking the phospholipase Cγ (PLCγ) signaling pathway with either a specific PLCγ inhibitor or knockdown PLCγ reduced the expression levels of both FUT9 and ST3Gal4 mRNAs and inhibited L1-mediated neuronal differentiation. These results demonstrate that L1 promotes neuronal differentiation from ESCs through the L1-mediated enhancement of FUT9 and ST3Gal4 expression.« less
  • Endothelial cells express two classical cadherins, VE-cadherin and N-cadherin. VE-cadherin is absolutely required for vascular morphogenesis, but N-cadherin is thought to participate in vessel stabilization by interacting with periendothelial cells during vessel formation. However, recent data suggest a more critical role for N-cadherin in endothelium that would regulate angiogenesis, in part by controlling VE-cadherin expression. In this study, we have assessed N-cadherin function in vascular development using an in vitro model derived from embryonic stem (ES) cell differentiation. We show that pluripotent ES cells genetically null for N-cadherin can differentiate normally into endothelial cells. In addition, sprouting angiogenesis was unaltered,more » suggesting that N-cadherin is not essential for the early events of angiogenesis. However, the lack of N-cadherin led to an impairment in pericyte covering of endothelial outgrowths. We conclude that N-cadherin is necessary neither for vasculogenesis nor proliferation and migration of endothelial cells but is required for the subsequent maturation of endothelial sprouts by interacting with pericytes.« less
  • The murine embryonic stem cell test (EST) is designed to evaluate developmental toxicity based on compound-induced inhibition of embryonic stem cell (ESC) differentiation into cardiomyocytes. The addition of transcriptomic evaluation within the EST may result in enhanced predictability and improved characterization of the applicability domain, therefore improving usage of the EST for regulatory testing strategies. Transcriptomic analyses assessing factors critical for risk assessment (i.e. dose) are needed to determine the value of transcriptomic evaluation in the EST. Here, using the developmentally toxic compound, flusilazole, we investigated the effect of compound concentration on gene expression regulation and toxicity prediction in ESCmore » differentiation cultures. Cultures were exposed for 24 h to multiple concentrations of flusilazole (0.54-54 {mu}M) and RNA was isolated. In addition, we sampled control cultures 0, 24, and 48 h to evaluate the transcriptomic status of the cultures across differentiation. Transcriptomic profiling identified a higher sensitivity of development-related processes as compared to cell division-related processes in flusilazole-exposed differentiation cultures. Furthermore, the sterol synthesis-related mode of action of flusilazole toxicity was detected. Principal component analysis using gene sets related to normal ESC differentiation was used to describe the dynamics of ESC differentiation, defined as the 'differentiation track'. The concentration-dependent effects on development were reflected in the significance of deviation of flusilazole-exposed cultures from this transcriptomic-based differentiation track. Thus, the detection of developmental toxicity in EST using transcriptomics was shown to be compound concentration-dependent. This study provides further insight into the possible application of transcriptomics in the EST as an improved alternative model system for developmental toxicity testing.« less